1. Complete integration of carbene-transfer chemistry into biosynthesis.
- Author
-
Huang, Jing, Quest, Andrew, Cruz-Morales, Pablo, Deng, Kai, Pereira, Jose Henrique, Van Cura, Devon, Kakumanu, Ramu, Baidoo, Edward E. K., Dan, Qingyun, Chen, Yan, Petzold, Christopher J., Northen, Trent R., Adams, Paul D., Clark, Douglas S., Balskus, Emily P., Hartwig, John F., Mukhopadhyay, Aindrila, and Keasling, Jay D.
- Abstract
Biosynthesis is an environmentally benign and renewable approach that can be used to produce a broad range of natural and, in some cases, new-to-nature products. However, biology lacks many of the reactions that are available to synthetic chemists, resulting in a narrower scope of accessible products when using biosynthesis rather than synthetic chemistry. A prime example of such chemistry is carbene-transfer reactions1. Although it was recently shown that carbene-transfer reactions can be performed in a cell and used for biosynthesis2,3, carbene donors and unnatural cofactors needed to be added exogenously and transported into cells to effect the desired reactions, precluding cost-effective scale-up of the biosynthesis process with these reactions. Here we report the access to a diazo ester carbene precursor by cellular metabolism and a microbial platform for introducing unnatural carbene-transfer reactions into biosynthesis. The α-diazoester azaserine was produced by expressing a biosynthetic gene cluster in Streptomyces albus. The intracellularly produced azaserine was used as a carbene donor to cyclopropanate another intracellularly produced molecule—styrene. The reaction was catalysed by engineered P450 mutants containing a native cofactor with excellent diastereoselectivity and a moderate yield. Our study establishes a scalable, microbial platform for conducting intracellular abiological carbene-transfer reactions to functionalize a range of natural and new-to-nature products and expands the scope of organic products that can be produced by cellular metabolism.The α-diazoester azaserine can be produced by Streptomyces albus engineered with a biosynthetic gene cluster and act as the carbene precursor for coupling with intracellularly produced styrene to generate unnatural amino acids containing a cyclopropyl group. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF